Induction of adipose-derived stem cell into motoneuron-like cells using selegiline as preinducer

Abstract

Cell therapy is one of the approaches taken to treatment of spinal cord disorders. In this study, adipose-derived stem cells (ADSCs) were induced to form motoneuron-like cells (MNLCs) using selegiline as preinducer, as well as Shh and all trans-retinoic acid (RA) as inducers. Selegiline was reported to induce the embryonic stem cells and bone marrow stromal cells into neuronal phenotype. ADSCs were evaluated using CD90, CD44, CD 49d, CD106, CD31, CD45, lipogenesis and osteogenesis. Dose response and time course studies were used in selecting the optimal concentration for selegiline using the percentage of viable cells (PVC) and percentages of immunoreactive cells (PIC) to nestin and neurofilament 68. Accordingly, such studies were used in selecting the optimal dose for RA using PVC and PIC to islet-1 and oligo-2. The expression of islet-1, oligo-2 and HLXB9 was evaluated using RT-PCR and immunocytochemistry. Real-time PCR was utilized in order to quantify the expression of islet-1, oligo-2 and HLXB9. ADSCs were immunoreactive to CD90, CD44 and CD 49d with consistent differentiation osteogenic and lipogenic cells. The optimal concentrations of selegiline and RA were 10−9mM and 2×10−8M, respectively. After two days, MNLCs showed high oligo-2 expression. MNLCs innervated myotubes; also, the release rate of synaptic vesicles using FM1-43 followed exponential decay model, and this rate in the induced MNLCs was approximately three times of that in the preinduced cells.